The present invention relates generally to an optical method and apparatus for diagnosing human spermatozoa and more particularly to an optical method and apparatus for determining the sperm count and sperm motility.
It is most important to be able to determine when a male is truly subfertile. Two parameters commonly used by urologists to measure fertility are sperm count and sperm motility. Determining sperm count and motility by the well known technique of observation through a microscope is time consuming and has not been entirely satisfactory to investigators for many years. More recently, absorption spectrophotometry, dye stained fluorometry, DNA determination and flow cytometry have been used to determine sperm count while absorption spectrophotometry, time-lapse photography, cinematography, and laser light-scattering have been used to determine sperm motility. In addition, multiple exposure photography (MEP) and computer based methods of bi-level picture processing (BPP) systems have been used for quantitative evaluation of the sperm density and motility.
For more than a half a century, fluorescence, absorption, and excitation spectroscopies have been extensively used as a probe to acquire fundamental information about the various physical, chemical, and biological processes in nature. Recently, the optical methods have been extended and applied to the medical field to diagnose disease. Extrinsic fluorescent dye markers have been used but interact with the native cellular environment. The majority of the molecular systems of the cells are made of proteins, nucleic acids and lipid, with fluorescing and non-fluorescing chromophores. These systems are known to predominantly luminesce in the UV and visible spectral regions. Recently, a new technique has been developed using the "intrinsic" nature fluorophors to characterize the physiological state of the abnormal systems as compared to the corresponding normal systems. These native fluorophors display well defined spectral features that characterize the local environment and state of the cells. Their spectroscopy is sensitive to the environment of pH, redox potential, bonding sites, polarity, ion concentration and so on. Flavins are known to fluorescence in the visible spectrum region from about 510 to 530 nm. and exhibit spectral changes in different environments. Flavins are located in mitochondria.
In U.S. Pat. No. 4,290,433 to R. R. Alfano there is disclosed a technique for measuring tooth decay using fluorescence spectroscopy. Differences in the fluorescence spectra of carious and noncarious regions of teeth are clearly detected and used to determine the presence of caries. This technique has since been extended to the diagnosis of cancer and to atherosclerosis.
The following is a list of known publications pertinent to this invention:
Udenfriend, S: Fluorescenced Assay in Biology and Medicine, Vol. 1. New York: Academic Press, 1962; Udenfriend, S: Fluorescence Assay in Biology and Medicine, Vol. 2. New York; Academic Press, 1969; Pringsheim, P: fluorescence and phospherescence. New York Interscience, 1949; Fasman, G. D. Ed., Handbook of biochemistry and molecular biology, 3rd. Ed., Cleveland, Ohio: CRC Press, 1975, pp. 205-210; Chance, B., B. Schoener: Fluorometric studies of flavin compounds of the respiratory chain in Flavins and flavoprotein, E. C. Slater, Ed. Amsterdam, The Netherlands: Elsevier, 1966; Alfano, R. R. S. S. Yao: Human teeth with and without caries studies by visible fluorescent spectroscopy. J. Dent. Res. 60: 120-122, 1981; Alfano R. R., D. Tata, J. Cordero, P. Tomashefsky, F. Longo, M. A. Alfano: Laser induced fluorescence spectroscopy from native cancerous and normal tissue. IEEE J.QE. QE-20: 1507-1511, 1984; Kittrel, C., R. L. Willet, C. de Los Santose Pacheo, N. B. Ratliff, Jr., R. Kramer, E. G. Malk, M. S. Feld: Diagnosis of fibrous arterial atherosclerosis using fluorescence. app. Opt. 24: 2280-2281, 1985. Mann, T., C Lutwak-Mann, Male Reproductive function and semen, Berlin, Germany, Springer-Verlag, 1981; Badenoch, D. F., C. G. Fowler, P. R. Evans, P. A. Lowrey: DNA content of human semen-an objective measurement of sperm density. British J. Urol. 57: 230-232, 1985; Garner, D. L., Gledhill, B. L., Pinkel, D., Lake, S., Stephenson, D., Van Dilla, M. A., Jhonnon, L. A.: Qualification of the X- and Y-Chromosome-bearing spermatozoa of domestic animals by flow cytometry. Biol. Repro. 28: 312-321, 1983; and Sokoloski, J. E., L. Blasco, B. T. Storey, D. P. Wolk: Turbidimetric analysis of human sperm motitlity. Fertil. steril. 28: 1337-1341, 1977.
It is an object of this invention to provide a new and improved method and apparatus for measuring sperm count in a specimen of semen.
It is another object of this invention to provide a new and improved method and apparatus for measuring sperm motility in a specimen of semen.